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Merck

H6380

Sigma-Aldrich

Hexokinase from Saccharomyces cerevisiae

lyophilized powder, ≥350 units/mg protein, Protein ≥10 % by biuret

Sinónimos:

ATP:D-Hexose-6-phosphotransferase, Hexokinase from yeast

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About This Item

Número de CAS:
Comisión internacional de enzimas:
Número CE:
Número MDL:
Código UNSPSC:
12352204
eCl@ss:
32160410
NACRES:
NA.54

origen biológico

Saccharomyces cerevisiae

Nivel de calidad

Formulario

lyophilized powder

actividad específica

≥350 units/mg protein

mol peso

dimer 110 kDa

composición

Protein, ≥10% biuret

fabricante / nombre comercial

Sigma-Aldrich

técnicas

activity assay: suitable

color

white

pH

6

Nº de acceso UniProt

aplicaciones

life science and biopharma

actividad extraña

L-Glutamic dehydrogenase ≤0.05%
Creatine phosphokinase, myokinase, 6-phosphogluconic dehydrogenase and alcohol dehydrogenase ≤0.001%
Glucose-6-phosphate dehydrogenase ≤0.005%
Glutathione reductase ≤0.005%
Phosphoglucose isomerase ≤0.002%

temp. de almacenamiento

2-8°C

Información sobre el gen

bakers yeast ... HXK1(850614) , HXK2(852639)

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Descripción general

Research area: Cell signalling. Hexokinase (HK) from yeast is a 486 amino-acid containing enzyme with a α/β fold. It comprises a large and small domain separated by an active site cleft. Conserved glycine residues are essential for ATP and glucose binding. The two isoforms are HKI and HKII encoded by HXK1 and HXK2 gene, respectively.

Aplicación

Hexokinase from Saccharomyces cerevisiae has been used:
  • to investigate its amyloid fibril forming tendency using biophysical methods
  • in tracking ATP depletion in peroxisomes and reticulocyte lysates in import assays
  • in the synthesis of Mg-ADP-actin from Mg-ATP actin monomers

Acciones bioquímicas o fisiológicas

Catalyzes the phosphorylation of D-hexose sugars at the C6 position utilizing ATP as a phosphate source.

The rate of phosphorylation varies with different hexoses (pH 7.5, 30 °C).
D-fructose KM: 0.33 mM
D-glucose KM: 0.12 mM
D-mannose KM: 0.05 mM

Yeast hexokinase exists as two similar isoforms, PI and PII (A and B), with isoelectric points of 5.25 and 4, respectively.

Molecular Weight: ~ 54 kDa (monomer)
~110 kDa (dimer)
Optimal pH: 7.5 to 9.0
Extinction Coefficient: E1% = 8.85 (PI) and 9.47 (PII) at 280 nm

Activators: Hexokinase requires Mg2+ ions (KM = 2.6 mM) for activity. Hexokinase is activated by catecholamines and related compounds.

Inhibitors: sorbose-1-phosphate, polyphosphates, 6-deoxy-6-fluoroglucose, 2-C-hydroxy-methylglucose, xylose, lyxose, and thiol reactive compounds (Hg2+ and 4-chloromercuribenzoate)
Hexokinase II is a crucial enzyme in glucose metabolism. Deletion of gene in S. cerevisiae in the presence of high glucose displays oxidative growth and results in the production of high biomass as well as shortened diauxic shift. Hexokinase II is also involved in glucose repression.

Definición de unidad

One unit will phosphorylate 1.0 μmole of D-glucose per min at pH 7.6 at 25 °C, unless otherwise indicated below.

Forma física

Lyophilized powder containing phosphate/citrate pH approx. 7.0

Otras notas

Predominantly the PII isoform, produced from an overexpressing yeast strain.

Código de clase de almacenamiento

11 - Combustible Solids

Clase de riesgo para el agua (WGK)

WGK 3

Punto de inflamabilidad (°F)

Not applicable

Punto de inflamabilidad (°C)

Not applicable

Equipo de protección personal

Eyeshields, Gloves, type N95 (US)


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Visite la Librería de documentos

The energetics of Pex5p-mediated peroxisomal protein import
Oliveira M, et al.
Test, 278(41), 39483-39488 (2003)
Mechanism of actin polymerization revealed by cryo-EM structures of actin filaments with three different bound nucleotides
Chou SZ and Pollard TD
Proceedings of the National Academy of Sciences of the USA, 116(10), 4265-4274 (2019)
Large proteins have a great tendency to aggregate but a low propensity to form amyloid fibrils
Ramshini H, et al.
Testing, 6(1), e16075-e16075 (2011)
J A Diderich et al.
Applied and environmental microbiology, 67(4), 1587-1593 (2001-04-03)
Hexokinase II is an enzyme central to glucose metabolism and glucose repression in the yeast Saccharomyces cerevisiae. Deletion of HXK2, the gene which encodes hexokinase II, dramatically changed the physiology of S. cerevisiae. The hxk2-null mutant strain displayed fully oxidative
The high resolution crystal structure of yeast hexokinase PII with the correct primary sequence provides new insights into its mechanism of action
Kuser PR, et al.
Test, 275(27), 20814-20821 (2000)

Protocolos

Measure hexokinase activity using a continuous spectrophotometric rate-determination assay at 340 nm, catalyzing D-hexose sugar phosphorylation using ATP.

Measure hexokinase activity using a continuous spectrophotometric rate-determination assay at 340 nm, catalyzing D-hexose sugar phosphorylation using ATP.

Measure hexokinase activity using a continuous spectrophotometric rate-determination assay at 340 nm, catalyzing D-hexose sugar phosphorylation using ATP.

Measure hexokinase activity using a continuous spectrophotometric rate-determination assay at 340 nm, catalyzing D-hexose sugar phosphorylation using ATP.

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